CN102579509B - Selenium-enriched marasmius androsaceus toadstool extract preparation and preparation method thereof - Google Patents
Selenium-enriched marasmius androsaceus toadstool extract preparation and preparation method thereof Download PDFInfo
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- CN102579509B CN102579509B CN201210014153.2A CN201210014153A CN102579509B CN 102579509 B CN102579509 B CN 102579509B CN 201210014153 A CN201210014153 A CN 201210014153A CN 102579509 B CN102579509 B CN 102579509B
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Abstract
The invention discloses a selenium-enriched marasmius androsaceus toadstool extract preparation which is prepared by the steps that: based on marasmius androsaceus toadstool as a carrier for enriching selenium element, a liquid submerged fermentation technology is adopted, 10-30mg/ml of sodium selenite (Na2SeO3.5H2O) is added into mycelium and metabolite of the marasmius androsaceus toadstool in the growth process according to the volume weight ratio, so that the selenium content in the cultured product of the marasmius androsaceus toadstool reaches 240-400mu g/g; and an excipient is added according to the conventional technology, so that the selenium-enriched marasmius androsaceus toadstool extract preparation is prepared. The selenium-enriched marasmius androsaceus toadstool extract preparation can be used as a medicine for treating multiple pain diseases such as nervous headache, pain in waist and lower extremities, sciatica and the like, and is remarkable in treatment effect; compared with the similar medicines, the selenium-enriched marasmius androsaceus toadstool extract preparation has the characteristics of being high in cure rate, free from side effect or addiction and the like; and the selenium-enriched marasmius androsaceus toadstool extract preparation is simple in production technique, convenient in operation and remarkable in selenium enrichment effect, and has potential application and development values.
Description
Technical field
The present invention relates to extract formulation of a kind of bacterium and preparation method thereof, be specifically related to rich selenium marasmius androsaceus extract formulation and preparation method thereof.
Background technology
Selenium is the trace element of needed by human, and selenium has the free radical of removing, anti-inflammatory, antitumor and lipotropism matter oxygenizement, also has and delays senility simultaneously, improves the functions such as immunity of organisms.In medicine and food, directly add inorganic selenium salt, biological activity is low, be not only difficult to absorb, and toxic side effect, result of use is undesirable.People mainly rely on food and obtain selenium at present, and biogenic organoselenium, compared with inorganic selenium, has absorption rate high, the advantages such as safety non-toxic.China has 72% area to belong to lack selenium area, and in diet formula, selenium content is on the low side, researches and develops rich selenium product, is to improve people constitution, the important channel that strengthens disease resistance.
Relevant expert points out, selenium can participate in the important Metabolic activity of organism, has Cell protection film, improves immunity and anticancer effect.The concentration that promotes selenium in blood can be treated tumour effectively, organoselenium albumen can not only kill tumour cell fast, dwindles focus, can also " target " kill the tumour cell being diffused in other internal organs, blood and lymph, stop the transfer and relapse of tumour, without any side effects.
Exploitation biogenic organoselenium, the mushroom that particularly bio-transformation ability is strong is the effective way that selenium source is provided to body.Different edible mushroomss are different to the tolerance of selenium.Same edible mushrooms, different selenium contents is different to mycelial growth effect, selenium concentration is too large, can suppress growth, and output is sharply declined, and makes mycelium in cessation of growth cessation state, even dead; Selenium concentration is too low, does not reach again the effect of rich selenium.Select the bacterial classification strong to selenium tolerance, selecting best rich selenium rate is one of problem of current Scientific Research Workers further investigation.
Marasmius androsaceus is a kind of medicine-food two-purpose bacterium of preciousness, the multiple neuropathic pain diseases such as sciatica, trigeminal neuralgia, migraine are had to good curative effect, along with the proof repeatedly of development and the clinical trial of modern science and technology, the intension that the pharmaceutical use of marasmius androsaceus is constantly made new advances by excavation.
Because the marasmius androsaceus speed of growth is slower, natural resource are limited, production unit adopts solid culture and liquid deep layer culture technique to produce marasmius androsaceus mycelium and nutrient solution more at present, in order to meet clinical needs, but it is also rare for the report of medicine, foodstuffs industry to produce marasmius androsaceus mycelium by this biotechnology of liquid submerged fermentation, confirm through clinical medical research, the effect of mycelium and sporophore is identical, can carry out by means of this biotechnology of liquid state fermentation the depth exploitation of marasmius androsaceus.
Through the sci-tech information institution world, Shanxi Province, state's interconnect and data of optical disk library searching, according to the data retrieving, still there is no the open report of the rich selenium research of marasmius androsaceus at present.
Summary of the invention
The object of this invention is to provide a kind of rich selenium marasmius androsaceus extract formulation and preparation method thereof, the object of the invention is to be achieved through the following technical solutions: be the carrier using marasmius androsaceus as Enriching Selenium element, adopt liquid submerged fermentation technology, obtain in the short period of time a large amount of mycelium and the meta-bolites of rich selenium, in order to produce the rich selenium marasmius androsaceus extract formulation that can treat nervous headache, pain in the waist and lower extremities.
Tolerance and the transformation efficiency of marasmius androsaceus to selenium is higher with respect to general bacterial classification.The present invention, from the bio-transformation capability analysis to selenium, selects the marasmius androsaceus that rich selenium rate is high (Marasmius androsaceus) as bacterial classification, culture presevation unit: Chinese common micro-organisms culture presevation administrative center.
The carrier of the present invention using marasmius androsaceus as Enriching Selenium element, bacterial strain code: 5.152, be purchased from Chinese common micro-organisms culture presevation administrative center, adopt liquid submerged fermentation technology, in the mycelium of marasmius androsaceus and meta-bolites process of growth, add Sodium Selenite (Na according to weight proportion
2seO
35H
2o), make selenium content in Marasmius androsaceus (L.ex Fr.) Fr. fungus extract reach 240～400 μ g/g, add vehicle according to common process, make rich selenium marasmius androsaceus extract formulation.
The preparation method of rich selenium marasmius androsaceus extract formulation of the present invention:
A, slant culture
Substratum: the Sodium Selenite (Na that adds 3～30 mg/100ml in PDA synthetic medium
2seO
35H
2o), make the selenium content in substratum reach 9～90 μ g/ml.
Culture condition: cultivate 9～11 days under 25 ℃ ± 1 ℃ constant temperature.
B, liquid shaking bottle seed culture:
Substratum: wheat bran 4～6%, glucose 2～4%, yeast extract paste 0.1～0.3%, KH
2pO
40.05～0.3%, MgSO
40.03～0.15%, the Sodium Selenite (Na of 3～30mg/100ml
2seO
35H
2o), moisturizing, to determining volume, makes the selenium content in substratum reach 9～90 μ g/ml; Wherein wheat bran boils to remove slag for 30 minutes and gets filtrate.
Culture condition: 750 ml shaking flask loading amount 200ml, 130～180 revs/min of rotating speeds, 25 ℃ ± 1 ℃ of temperature is cultivated 13～15 days on rotary shaking table.
C, liquid shaking bottle fermentation culture:
Substratum: glucose 2～4%, corn steep liquor 0.3～1.0%, yeast powder 0.2～0.6%, KH
2pO
40.05～0.3%, MgSO
40.03～0.15%, the Sodium Selenite (Na of 3～30mg/100ml
2seO
35H
2o), moisturizing, to determining volume, makes the selenium content in substratum reach 9～90 μ g/ml.
Culture condition: 750 ml shaking flask loading amount 200ml, 130～180 revs/min of rotating speeds, 25 ℃ ± 1 ℃ of temperature is cultivated 8～9 days on rotary shaking table.
D, broth extraction process:
Leaching process by mycelia, bacterium liquid separate, bacterium liquid adopts rotatory evaporator to be condensed into medicinal extract, mycelia loft drier is dry, dried rich selenium marasmius androsaceus mycelia alcohol reflux, extracting solution concentrated extract mixes with bacterium immersion cream, and in mixed medicinal extract, selenium content reaches 240～400 μ g/g; Add vehicle according to common process, make rich selenium marasmius androsaceus tablet and other preparation.
Adopt preparation method of the present invention, the rich selenium rate of marasmius androsaceus mycelia can bring up to 13～15%, available data has no report, the present invention adopts advanced alcohol reflux production technique, make the effective ingredient polysaccharide containing in product, N.F,USP MANNITOL and trace element-selenium, can more intactly preserve, play the effect of anti-pain, the N.F,USP MANNITOL that said preparation contains can be removed oxyradical, reduce algogenic substance, activity that can also inhibition of pain endogenous neurotransmitter, or change the positively charged ion of induced pain and some protein with the avidity of negatively charged ion, change the membrane potential of susceptor, effect maintains more than 6 hours, and without knock-on rise phenomenon, therefore can make the headache that continues outbreak is effectively alleviated.The water extracting alcohol hypostasis ANA of marasmius androsaceus can significantly improve the threshold of pain.
The present invention is the medicine as multiple pain diseases such as treatment nervous headache, pain in the waist and lower extremities, sciatica with rich selenium marasmius androsaceus extract formulation, evident in efficacy, compared with similar drugs, has curative ratio high, has no side effect, without features such as habituation.Its production technique is simple, easy to operate, and rich selenium effect is remarkable, has potential application and exploitation value.
Embodiment
The marasmius androsaceus that the present invention selects, strain number: 5.152, introduce from Chinese common micro-organisms culture presevation administrative center.Adopt three grades of submerged fermentations to cultivate, fermenting process adopts aseptic technique, bacterium liquid after leaching process employing mycelia, bacterium liquid separate, separate concentrates, mycelia is dry, dried rich selenium marasmius androsaceus mycelia alcohol reflux, extracting solution concentrated extract mixes with bacterium immersion cream, and in mixed medicinal extract, selenium content reaches 240～400 μ g/g; Add vehicle according to common process, make rich selenium marasmius androsaceus tablet and other preparation.
The preparation method of rich selenium Marasmius androsaceus (L.ex Fr.) Fr. fungus extract is:
A, slant culture
Substratum: the Sodium Selenite (Na of PDA synthetic medium+10mg/100ml
2seO
35H
2o), make the selenium content in substratum reach 30 μ g/m l.
Culture condition: cultivate 9～11 days under 25 ℃ ± 1 ℃ constant temperature.
B, liquid shaking bottle seed culture:
Substratum: wheat bran 5.0%, glucose 3.0%, yeast extract paste 0.2%, KH
2pO
40.1%, MgSO
40.05% and the Sodium Selenite (Na of 10mg/100ml
2seO
35H
2o), moisturizing, to determining volume, makes the selenium content in substratum reach 30 μ g/ml; Wherein wheat bran boils to remove slag for 30 minutes and gets filtrate.
Culture condition: 750 ml shaking flask loading amount 200ml, 135 revs/min of rotating speeds, 25 ℃ ± 1 ℃ of temperature is cultivated 13 days on rotary shaking table.
C, liquid shaking bottle fermentation culture:
Substratum: glucose 3.0%, corn steep liquor 0.5%, yeast powder 0.4%, KH
2pO
40.1%, MgSO
40.05%, the Sodium Selenite (Na of 10mg/100ml
2seO
35H
2o), moisturizing, to determining volume, makes the selenium content in substratum reach 30 μ g/ml.
Culture condition: 750 ml shaking flask loading amount 200ml, 135 revs/min of rotating speeds, 25 ℃ ± 1 ℃ of temperature is cultivated 8～9 days on rotary shaking table.
D, broth extraction process:
Leaching process by mycelia, bacterium liquid separate, bacterium liquid adopts that rotatory evaporator is concentrated, mycelia loft drier is dry, dried rich selenium marasmius androsaceus mycelia alcohol reflux, extracting solution concentrated extract mixes with bacterium immersion cream, and in mixed medicinal extract, selenium content reaches 240～400 μ g/g; Add vehicle according to common process, make rich selenium marasmius androsaceus tablet and other preparation.
According to preparation method of the present invention, all right granulation agent of rich selenium Marasmius androsaceus (L.ex Fr.) Fr. fungus extract, capsule, oral liquor and other types of formulations.
Rich selenium marasmius androsaceus tablet in treatment angioneurotic headache for the present invention.Through Huazhou, Guangdong Province the People's Hospital clinical observation on the therapeutic effect 168 examples, treatment group is taken rich selenium marasmius androsaceus tablet " rich selenium anluotone tablet " (Shanxi Kangxin Pharmaceutical Industry Co., Ltd.'s production), control group is taken the conventional nonsteroidal antipyretic and analgesic such as amidopyrine compound, antondin, Ibuprofen BP/EP, is below the clinical observation experiment report of " rich selenium anluotone tablet " treatment angioneurotic headache.
Clinical efficacy comparison: treatment group: 84 examples, effective 68 examples (80.95%), effective 10 examples (11.9%), invalid 6 examples (7.14%), total effective rate 92.9%.
Control group: 84 examples, effective 55 examples (65.47%), effective 11 examples (13.09%), invalid 17 examples (20.23%), total effective rate 78.56%.
Take " rich selenium anluotone tablet " treatment angioneurotic headache, clinical obvious effective rate and total effective rate are all significantly higher than control group, and statistical study shows (P < 0.05), and treatment group is better than control group.
Through the clinical observation to 168 routine angioneurotic headache patients, total effective rate reaches more than 92%, rear respond well.Result shows: " rich selenium anluotone tablet " treatment angioneurotic headache is used the conventional nonsteroidal antipyretic and analgesic better efficacy such as amidopyrine compound, antondin, Ibuprofen BP/EP.It is slow that " rich selenium anluotone tablet " has onset, the length of holding time, and again without the advantage of habituation and other untoward reaction, long-term taking preferably.After taking medicine and coming into force, pain is that level is chronic to be alleviated until disappear, and is a kind of more satisfactory analgesia Chinese patent medicine, is worthy to be popularized.
" rich selenium anluotone tablet " treatment pain in the waist and lower extremities for the present invention, through Dianbai County the People's Hospital, pain in the waist and lower extremities patient clinical is observed to 160 examples, treatment group oral rich selenium anluotone tablet (Shanxi Kangxin Pharmaceutical Industry Co., Ltd.'s production) simultaneously on the basis for the treatment of of control group, 3 times/day, 1 time 1.See the observation of curative effect of " rich selenium anluotone tablet " treatment pain in the waist and lower extremities.
Clinical efficacy comparison: control group is used: 5% glucose injection 250 ml add compound injection of red sage root 20 ml intravenous drips, 1 times/day; 20% formula mannitol injection liquid 250 ml intravenous drips,, 1 times/day, stop using for 3 days afterwards by 120/min; Acute phase, adds Dexamethasone Injection 10 mg, and 1 times/day, after 3～5 days, decrement is stopped using.
Treatment group 80 examples, cure 41 examples, obvious 21 examples, and 12 examples that take a turn for the better, invalid 6 examples, total effective rate is 92.48%; Control group: 80 examples, cure 12 examples, obvious 24 examples, 21 examples that take a turn for the better, invalid 23 examples, total effective rate is 71.25%, through chi square test, X
2=11.53, P < 0.05, treatment group and control group Different therapeutical effect have significance, and treatment group is better than control group.
" rich selenium anluotone tablet " adopts the mycelium in extraction using alcohol marasmius androsaceus, and its chemical composition has N.F,USP MANNITOL, Rhabdasterol acetate, amino acid, p-Coumaric Acid, ergosterol, selenium element etc.The material in marasmius androsaceus with analgesic activity has p-Coumaric Acid, 30 carbonic acid, ergosterol, terpene lactones compound and saponin etc., and the trace element-selenium in bacterium can be removed interior free yl, get rid of vivotoxin, anti-oxidant, the generation that can effectively suppress lipid peroxide, prevent blood clot, remove cholesterol, strengthening immune function of human body, is a kind of more satisfactory analgesia Chinese patent medicine, without the advantage of habituation and other untoward reaction.
" rich selenium anluotone tablet " adopts selenium-enriched liquid submerged fermentation technology, extract effective constituent polysaccharide, N.F,USP MANNITOL in marasmius androsaceus, p-Coumaric Acid, selenium, by improving the ability of antioxidant radical of body, regulate body's immunity, thus the anti-pain effect of enhancing body." rich selenium anluotone tablet " easy administration, mouthfeel are good, not yet find that there is at present untoward reaction and habituation, and suitable long-term taking is the ideal medicament for the treatment of nervous headache, pain in the waist and lower extremities etc.Experiment shows, " rich selenium anluotone tablet " easy administration, and good effect, safe, suitable long-term taking, clinic is promoted the use of.
The above; only for preferably embodiment of the present invention, but protection scope of the present invention is not limited to this, is anyly familiar with in technical scope that those skilled in the art disclose in the present invention; the variation that can expect easily or replacement, within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain of claims.
Claims (3)
1. a preparation method for rich selenium marasmius androsaceus extract formulation, is characterized in that the carrier using marasmius androsaceus as Enriching Selenium element, bacterial strain code: 5.152, be purchased from Chinese common micro-organisms culture presevation administrative center; In the mycelium of marasmius androsaceus and meta-bolites process of growth, add Sodium Selenite according to weight proportion, make the selenium content in substratum reach 9～90 μ g/ml, carry out liquid submerged fermentation cultivation, finally fermented liquid extracted, specifically formed by following steps:
A, slant culture
Substratum: the Na of PDA synthetic medium+3～30 mg/100ml
2seO
35H
2o, makes the selenium content in substratum reach 9～90 μ g/ml,
Culture condition: cultivate 9～11 days under 25 ℃ ± 1 ℃ constant temperature;
B, liquid shaking bottle seed culture:
Substratum: wheat bran 4～6%, glucose 2～4%, yeast extract paste 0.1～0.3%, KH
2pO
40.05～0.3%, MgSO
40.03～0.15%, Na
2seO
35H
2o 3～30mg/100ml, moisturizing, to determining volume, makes the selenium content in substratum reach 9～90 μ g/ml, and wherein wheat bran boils to remove slag for 30 minutes and gets filtrate,
Culture condition: 750 ml shaking flask loading amount 200ml, 130～180 revs/min of rotating speeds, 25 ℃ ± 1 ℃ of temperature is cultivated 13～15 days on rotary shaking table;
C, liquid shaking bottle fermentation culture:
Substratum: glucose 2～4%, corn steep liquor 0.3～1.0%, yeast powder 0.2～0.6%, KH
2pO
40.05～0.3%, MgSO
40.03～0.15%, Sodium Selenite Na
2seO
35H
2o 3～30mg/100ml, moisturizing, to determining volume, makes the selenium content in substratum reach 9～90 μ g/ml,
Culture condition: 750 ml shaking flask loading amount 200ml, 130～180 revs/min of rotating speeds, 25 ℃ ± 1 ℃ of temperature is cultivated 8～9 days on rotary shaking table;
D, broth extraction process:
Mycelia, bacterium liquid are separated, bacterium liquid adopts rotatory evaporator concentrated, and mycelia is dry with loft drier, dried rich selenium marasmius androsaceus mycelia alcohol reflux, extracting solution concentrated extract mixes with bacterium immersion cream, and in mixed medicinal extract, selenium content reaches 240～400 μ g/g; Add vehicle according to common process, make rich selenium marasmius androsaceus extract formulation.
2. the rich selenium marasmius androsaceus extract formulation that prepared by preparation method according to claim 1.
3. rich selenium marasmius androsaceus extract formulation according to claim 2, is characterized in that: the formulation of described preparation is tablet, granule, capsule or oral liquid.
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| CN104073531B (en) * | 2014-07-15 | 2016-08-24 | 江苏神华药业有限公司 | A kind of deep layer liquid fermentation produces the method for Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide |
| CN104857027B (en) * | 2015-04-19 | 2019-01-18 | 吉林大学 | CNNS Marasmius T08 is preparing the medical application in analgesic |
| CN105838623A (en) * | 2016-04-15 | 2016-08-10 | 山西康欣药业有限公司 | Preparation method of selenium-rich Armillaria mellea powder |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1297740A (en) * | 2000-11-22 | 2001-06-06 | 张义元 | Compound medicine for migraine |
| CN1424054A (en) * | 2002-12-23 | 2003-06-18 | 上海本草生物医学工程研究所 | Bacterial Chinese medical preparation and preparing method thereof |
| CN1428334A (en) * | 2001-12-27 | 2003-07-09 | 湖南隆来福生物工程有限公司 | Alkaloid compound, pain-stopping medicine and its preparation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1297740A (en) * | 2000-11-22 | 2001-06-06 | 张义元 | Compound medicine for migraine |
| CN1428334A (en) * | 2001-12-27 | 2003-07-09 | 湖南隆来福生物工程有限公司 | Alkaloid compound, pain-stopping medicine and its preparation method |
| CN1424054A (en) * | 2002-12-23 | 2003-06-18 | 上海本草生物医学工程研究所 | Bacterial Chinese medical preparation and preparing method thereof |
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